Diprenylated flavonoids from licorice induce death of SW480 colorectal cancer cells by promoting autophagy: Activities of lupalbigenin and 6,8-diprenylgenistein.

ETHNOPHARMACOLOGICAL RELEVANCE: Licorice is a well-known herbal medicine, and we previously found that several licorice prenylated flavonoids could cause death of SW480 colorectal cancer cells by promoting autophagy. Given many kinds of prenylated flavonoids in licorice, the activities of other compounds deserve further investigation. In addition, the contribution of isoprenyl groups on the autophagy promotion activities has not been clarified. AIM OF THE STUDY: This study aimed to investigate whether lupalbigenin (LPB) and 6,8-diprenylgenistein (DPG), two licorice diprenylated flavonoids, could induce autophagic cell death of SW480 cells, and clarify the contribution of isoprenyl groups. MATERIALS AND METHODS: Cytotoxic activities of LPB and DPG were tested by using an MTT method, and apoptosis induction effects were evaluated by PI-Annexin V staining-based flow cytometry and protein levels of caspase-3 and PARP-1. Autophagy promotion effects of LPB and DPG were assessed by protein levels of LC3, p62, Akt and mTOR as well as number of autophagosomes in cells, and autophagy inhibitor chloroquine (CQ) was involved to identify the role of autophagy on LPB or DPG-caused death of SW480 cells. In addition, two groups of structurally similar diprenylated, mono-prenylated and free flavonoids were obtained from licorice, which were used to investigate the contribution of isoprenyl groups on their autophagy promotion activities. RESULTS: Both LPB and DPG significantly induced apoptosis of SW480 cells with strong cytotoxic activities, and meanwhile, they also promoted autophagy probably through the Akt/mTOR signaling pathway. Further studies indicated that LPB and DPG could induce autophagic cell death of SW480 cells. Moreover, isoprenyl groups contributed mainly to the cytotoxic and autophagy promotion activities of licorice prenylated flavonoids. CONCLUSION: This study reported for the first time that licorice diprenylated flavonoids LPB and DPG induced death of SW480 cells by promoting autophagy, which was mainly attributed to the isoprenyl groups. The results provided theoretical basis for researches on anti-colorectal cancer drugs and their structural modification.

Prenylated phenolic compounds from licorice (Glycyrrhiza uralensis) and their anti-inflammatory activity against osteoarthritis.

Osteoarthritis is a significant driver of disability in the elderly with increasing prevalence, and inflammation plays a vital role on its etiology. Licorice is commonly used as a traditional Chinese medicine or food additive, and its prenylated phenolic compounds were recently reported to be able to inhibit osteoarthritis with anti-inflammatory activity. In order to explore more anti-osteoarthritic prenylated phenolic compounds from licorice, we isolated ten compounds (1-10), with three new ones (1-3), from the ethyl acetate extract of Glycyrrhiza uralensis. Compound 2 (glycyuralin R) was a racemic 3-phenoxy-chromanone, and we achieved its chiral separation for the first time. Compounds 1, 2, 7 and 8 showed significant NO inhibitory ability in IL-1ß-stimulated mouse primary chondrocytes, and we further confirmed the anti-inflammatory activity of 1 (glycyuralin Q) by evaluating its effect on osteoarthritis-related iNOS, COX-2, TNF-α, IL-6, MMP3, MMP13 and NF-κB based on various experimental methods. These results clarified the potential of several prenylated phenolic compounds, especially 1 in licorice, as the lead compounds for osteoarthritis.

Morus alba leaves ethanol extract protects pancreatic islet cells against dysfunction and death by inducing autophagy in type 2 diabetes.

BACKGROUND: Protection of pancreatic islet cells against dysfunction or death by regulating autophagy is considered to be an effective method for treatment of type 2 diabetes mellitus (T2DM). Morus alba leaves (mulberry leaves), a popular herbal medicine, have been used for prevention of T2DM since ancient times. PURPOSE: This study aimed to clarify whether Morus alba leaves ethanol extract (MLE) could protect islet cells in vivo and in vitro by regulating autophagy in T2DM, and explore the possible mechanism of action. METHODS: The main chemical constituents in MLE were analyzed by HPLC. The T2DM rat model was induced via high-fat diet combined with peritoneal injection of low-dose streptozotocin, and MLE was administered by oral gavage. Fasting blood glucose (FBG) and plasma insulin were measured, and homeostatic model assessment of ß cell function (HOMA-ß) and insulin resistance (HOMA-IR) were determined. The histomorphology of pancreas islets was evaluated by haematoxylin and eosin staining. In palmitic acid (PA)-stressed INS-1 rat insulinoma cells, cell viability was assayed by an MTT method. Expression of the autophagy-related proteins LC3 I/II, p62, p-AMPK and p-mTOR in islet tissues and INS-1 cells was evaluated by western blotting or immunohistochemistry analysis. RESULTS: The four main chemical constituents in MLE were identified as chlorogenic acid, rutin, isoquercitrin and quercitrin. MLE ameliorated hyperglycemia, insulin resistance and dyslipidemia of T2DM rats with prominent therapeutic effect. Further study indicated that MLE observably improved islet function, alleviated islet injury of T2DM rats, and inhibited PA-induced INS-1 cell death. On the other hand, MLE significantly induced autophagy in islet cells both in vivo and in vitro, and autophagy inhibitors abolished its therapeutic effect on T2DM rats and protective effect on islet cells. Apart from this, MLE markedly activated the AMPK/mTOR pathway in INS-1 cells, and the AMPK inhibitor prevented the autophagy induction ability of MLE. CONCLUSION: Together, MLE could protect islet cells against dysfunction and death by inducing AMPK/mTOR-mediated autophagy in T2DM, and these findings provide a new perspective for understanding the treatment mechanism of Morus alba leaves against T2DM.

Discussion of Correlation Between Chronic Psychological Stress Induced Liver Depression and Spleen Deficiency Syndrome and Microbiota-gut-brain Axis and Study on Intervention Mechanism of Xiaoyaosan / 中国实验方剂学杂志

Psychological stress （pressure） has gradually become an important factor affecting human physical and mental health， and is one of the important factors in the progression of clinical chronic refractory diseases. Psychological stress response can be attributed to the category of emotional illness in traditional Chinese medicine （TCM）. In the theory of TCM， the liver controls dispersion and regulates mental activity. Relevant scholars believe that the liver is the core of psychological stress response in the TCM theory. When being stimulated by chronic or repeated psychological stress， the body can gradually change from the initial depression and anxiety-rela behaviors to the gastrointestinal dysfunctions， which is similar to the formation process of the syndrome of liver depression and spleen deficiency with liver-spleen disharmony. The syndrome of liver depression and spleen deficiency is also the common syndrome of TCM for psychological stress-related diseases. With the effect in soothing liver and invigorating spleen， Xiaoyaosan has been regarded as the classic formula for anti-stress. Modern studies have showed that gut microbiota not only get involved in the movement， structure and function of the digestive tract， but also affect the brain function and behavior of the host through the gut-brain axis. Therefore， the microbiota-gut-brain axis has become an important part of psychological stress to trigger the body's gastrointestinal symptoms and abnormal brain behaviors. Focusing on psychological stress， the authors explored the correlation between the syndrome of liver stagnation and spleen deficiency and the microbiota-gut-brain axis， and the intervention mechanism of Xiaoyaosan， so as to enrich the scientific connotation of the syndrome of liver stagnation and spleen deficiency.

Ultrastructure Characteristics of Different Chinese Medicine Syndromes of Helicobacter pylori-Correlated Gastric Diseases.

OBJECTIVE: To explore the ultrastructure characteristics of patients with dampness-heat of Pi (Spleen)-Wei (Stomach) syndrome (DHPW) and Pi-qi deficiency syndrome (PQD), both of which are Helicobacter pylori (Hp)-correlated gastric diseases (HPCG), and implicate a helpful hint for the clinical microcosmic syndrome differentiation. METHODS: Fourteen gastric mucosa samples from 6 chronic gastritis (CG) and 6 active peptic ulcer (including 8 DHPW, 4 PQD) as well as 2 healthy volunteers were collected and tested for Hp infection. The ultrastructure of gastric mucosa was observed under the transmission electron microscope (TEM). RESULTS: Among 14 gastric mucosa samples, 8 of them were Hp positive (6 DHPW and 2 PQD), which were all accordance with the results screened by supermicro-pathological method. Under TEM, the normal gastric mucosa, with tidy microvilli and abundant in mucus granules, mitochondria and rough endoplasmic reticulum distributed evenly, and with smooth nucleus membrane. But in those specimens of DHPW with Hp infection, microvilli were presented with burr shape. Especially, those samples from dampness-heat syndrome with predominant heat type (DHSH) patients were more obvious, with microvilli damaged, mitochondria concentrated and distributed in disorder, secretory tubule extended. In dampness-heat syndrome with predominant dampness type (DHSD) patients, mucus granules aggregated obviously, mitochondria swelled and blurred, and rough endoplasmic reticulum crowded. For 2 samples of DHPW without Hp infection, their microvilli were intact, with mitochondria increased and gathered but well-distributed, and secretory tubule extended mildly. In 2 PQD patients with Hp positive, the specimens of microvilli were sparse, and their mucus granules and mitochondria were decreased, with fractured crests and vacuole, secretory tubules extension to nucleus membrane, and rough endoplasmic reticulum extension in a pool-like way, and nucleus condensed. The 2 samples from PQD patients without Hp infection were characterized with intact microvilli, decreased mitochondria, fractured crest and extended rough endoplasmic reticulum in a pool-like way. CONCLUSION: It's obviously different in ultrastructure of DHPW and PQD patients under TEM, which may give a helpful hint for the microcosmic syndrome differentiation of HPCG.

Segmented scan modes and polarity-based LC-MS for pharmacokinetic interaction study between Fufang Danshen Dripping Pill and Clopidogrel Bisulfate Tablet.

Fufang Danshen Dripping Pill (FDDP) and Clopidogrel Bisulfate Tablet (CBT) are usually combined for treatment of coronary artery diseases in clinical. To investigate the pharmacokinetic interaction between FDDP and CBT after oral administration of FDDP, CBT and their combination in rats, a novel LC-MS method with segmented scan modes (multiple reaction monitoring and selected ion monitoring) and polarity (positive and negative ionization) was developed. Clopidogrel and the main active ingredients of FDDP, with different chemical and ionization properties, were simultaneously quantified in plasma in a single run. The method was validated in terms of specificity, linearity, precision, accuracy, recovery, matrix effect and stability. As a result, co-administration of FDDP and CBT significantly altered the pharmacokinetic parameters of danshensu, ginsenoside Rb1, dihydrotanshinone I, tanshinone I and tanshinone IIA of FDDP, as well as clopidogrel. Mechanism studies suggested that induction of liver cytochrome P450 isozymes CYP2C11 and CYP3A1 by co-administration, as well as inhibition of carboxyl esterase 1, was partly responsible for FDDP-CBT pharmacokinetic interactions. The developed LC-MS method could be used to simultaneously quantify different types of in vivo analytes in a single run, and the results could be used for clinical medication guidance of FDDP and CBT.

[Expressions of MMP-7 and TIMP-1 in Chronic Stomach Disease Patients with Different Syndromes of Chinese Medicine].

Objective To observe the expressions of matrix metalloproteinase-7 (MMP-7) and tissue inhibitor of metalloproteinase-1 (TIMP-1 ) in chronic stomach disease patients with different syn- dromes of Chinese medicine (CM) , and their relationships with Helicobacter pylori ( Hp ) infection. Meth- ods Totally 117 chronic stomach disease patients were recruited, and 11 healthy volunteers were also recruited. Chronic stomach disease patients were assigned to Pi-Wei dampness-heat syndrome (PWDHS, 57 cases) , disharmony of Gan and Wei syndrome (DGWS, 30 cases) , and Pi qi deficiency syndrome (PQDS, 30 cases) by syndrome typing. Healthy volunteers were recruited as the healthy con- trol group. Hp infection was detected using methylene blue dyeing and rapid urease test (RUT). The degree of inflammation was observed by conventional HE staining. The protein expressions of MMP-7 and TIMP-1 were detected qualitatively and positioningly using immunohistochemical method. Results Patients with PWDHS and patients with DGWS had equivalent Hp infection rate and degree. They showed a slightly increasing tendency than patients with PQDS, but with no statistical difference (P >0. 05). Com- pared with PWDHS and PQDS groups, more severe inflammation of mucosa occurred in patients with DG- WS (P <0. 05). More severe inflammation of mucosa occurred in patients with PWDHS than in those with PQDS, but with no statistical difference (P >0. 05). The severity of gastric mucosal inflammatory activity was sequenced from high to low as PWDHS, DGWS, PQDS, all with statistical difference (P <0. 05). Compared with Hp negative patients, the gastric mucosal inflammatory activity was more severe in Hp positive patients with PWDHS, DGWS, PQDS. The gastric mucosal inflammatory activity was more se- vere in Hp positive patients with PWDHS and PQDS (P <0. 05). Compared with the healthy control group, the expression level of TIMP-1 in gastric mucosa increased in patients with PWDHS, DGWS, PQDS (P <0. 05, P <0. 01) ; the expression level of MMP-7 increased in Hp negative patients with PWDHS (P < 0. 05). Compared with Hp negative patients with PWDHS, the expression level of MMP-7 decreased in Hp positive patients with PWDHS (P <0. 05). Compared with the PQDS group, the expression level of TIMP-1 decreased in the PWDHS group (P <0. 01). The severity of gastric mucosal inflammation was negatively correlated with the expression level of MMP-7, and positively correlated with the expression level of TIMP- 1 (P <0. 01). Hp infection degree was not obviously correlated with the expression level of MMP-7 in gastric mucosa (P >0. 05) , but positively correlated with the expression level of TIMP-1 in gastric mucosa (P <0. 05). Of them, the expression level of MMP-7 in gastric mucosa was positively correlated with the expression level of TIMP-1 in gastric mucosa (P <0. 01). Conclusions Comparatively lower expression of MMP-7 in gastric mucosal inflammation and imbalanced expression of TIMP-1 might be two of the pathogeneses of chronic stomach disease. Their various expressions in different CM syndromes might have certain expositions for microscopic research on "different syndromes of the same disease". Emotional fluctuation might also be one of important factors for chronic stomach disease.

Efficacy of crest herbal toothpaste in "clearing internal heat": a randomized, double-blind clinical study.

Objective. Evaluation of the efficacy of Crest Herbal Crystal Toothpaste in "clearing internal heat." Methods. This was a randomized, double-blind, controlled parallel design clinical test of a product that was already on the market. 72 subjects were randomly assigned to control group (group A with Colgate Herbal Salty Toothpaste) or treatment group (group B with Crest Herbal Crystal Toothpaste) with ratio of 1 : 2. Subjects were instructed to brush with 1g toothpaste for 2 minutes each time, 2 times per day in a 4-aweek test period; measurement with the rating scale on the efficacy of "clearing internal heat" for the herbal toothpaste was done at baseline, 2 weeks, and 4 weeks of toothpaste usage. Results. The rating scale on efficacy of "clearing internal heat" for the herbal toothpaste reveals that the primitive points of 72-case intention-to-treat (ITT) analysis and 67-case per-protocol (PP) analysis for subjects in group A and subjects in group B were found to be reduced progressively with statistical significance (P < 0.05). The overall effective rates for group A and group B were, respectively, 62.50%, 56.25% (ITT) and 62.50%, 60.64% (PP). The statistical results indicated that the symptoms of fire-heat for both groups of subjects have been improved after application of toothpaste. Conclusion. The efficacy of Crest Herbal Crystal Toothpaste in "clearing internal heat" was confirmed by the trial as compared to Colgate Herbal Salty Toothpaste. And its efficacy was objectively evaluated by the rating scale on efficacy of "clearing internal heat."

Metabonomics-based study of clinical urine samples in suboptimal health with different syndromes.

Objective. To explore the urinary biochemistry features of syndromes of traditional Chinese medicine (TCM) such as syndrome of stagnation of liver Qi, spleen deficiency, liver Qi stagnation, and spleen deficiency (LSSDS) in sub-optimal health status (SHS). Methods. 12 cases for each syndrome group in SHS were selected, 12 subjects were used as a normal control group, and (1)H NMR detection was, respectively, carried out, and the data was corrected by the orthogonal signal correction (OSC) and then adopted a partial least squares (PLS) method for discriminate analysis. Results. The OSC-PLS (ctr) analysis results of the nuclear overhauser enhancement spectroscopy (NOESY) detection indicated that the syndromes in SHS could be differentiated, and there were significant differences in the levels of metabolites of the urine samples of the four groups; the biomarkers of LSSDS in SHS were found out. The contents of citric acid (2.54 and 2.66), trimethylamineoxide (3.26), and hippuric acid (3.98, 7.54, 7.58, 7.62, 7.66, 7.82, and 7.86) in the urine samples of LSSDS group were lower than that of the normal control group. Conclusion. There are differences in the (1)H-NMR metabolic spectrum of the urine samples of the four groups, and the specific metabolic products of the LSSDS in SHS can be identified from metabonomics analysis.

Development and evaluation of Chinese medicine fire-heat syndrome scale in oral cavity for measuring Chinese herb toothpaste.

OBJECTIVE: To formulate the standard measuring tool for the evaluations on fire-heat syndrome in oral cavity by means of Chinese medicine (CM). METHODS: The measuring scale for fire-heat syndrome in the oral cavity by means of CM was investigated by symptom collection, item pool formulation, item selection, pre-investigation, evaluations on the reliability, validity and reactivity of the measuring scale, according to the principles for measuring scale design and under the guidance of CM theories. RESULTS: The measuring scale was composed of two integrative parts: the self-filling section and the interview section. As far as the reliability was concerned, the total Cronbach α coefficient of the measuring scale was 0.866, the total test-retest reliability coefficient was 0.726 and the split-half reliability coefficient was 0.851. As far as the validity was concerned, the scores for the subjects of fire-heat syndrome in oral cavity and healthy people in their oral cavity in the items of symptoms were statistically different (P<0.01); three common divisors were extracted according to the theoretical dimensions, the accumulated contribution rate was 63.468%. As far as the reactivity was concerned, the difference between the symptom scores before and after the test in which 31 subjects used the Chinese herb toothpaste was statistically significant (P<0.01). CONCLUSIONS: This measuring scale has relatively good reliability, validity and reactivity, and it can be used in an objective quantitative evaluation on patients suffering from fire-heat syndrome in oral cavity, and thus lay the foundations for the evaluations on the therapeutic effects of Chinese herb toothpaste on fire-heat syndrome in oral cavity.

Development and evaluation of Chinese medicine fire-heat syndrome scale in oral cavity for measuring Chinese herb toothpaste / 中国结合医学杂志

<p><b>OBJECTIVE</b>To formulate the standard measuring tool for the evaluations on fire-heat syndrome in oral cavity by means of Chinese medicine (CM).</p><p><b>METHODS</b>The measuring scale for fire-heat syndrome in the oral cavity by means of CM was investigated by symptom collection, item pool formulation, item selection, pre-investigation, evaluations on the reliability, validity and reactivity of the measuring scale, according to the principles for measuring scale design and under the guidance of CM theories.</p><p><b>RESULTS</b>The measuring scale was composed of two integrative parts: the self-filling section and the interview section. As far as the reliability was concerned, the total Cronbach α coefficient of the measuring scale was 0.866, the total test-retest reliability coefficient was 0.726 and the split-half reliability coefficient was 0.851. As far as the validity was concerned, the scores for the subjects of fire-heat syndrome in oral cavity and healthy people in their oral cavity in the items of symptoms were statistically different (P<0.01); three common divisors were extracted according to the theoretical dimensions, the accumulated contribution rate was 63.468%. As far as the reactivity was concerned, the difference between the symptom scores before and after the test in which 31 subjects used the Chinese herb toothpaste was statistically significant (P<0.01).</p><p><b>CONCLUSIONS</b>This measuring scale has relatively good reliability, validity and reactivity, and it can be used in an objective quantitative evaluation on patients suffering from fire-heat syndrome in oral cavity, and thus lay the foundations for the evaluations on the therapeutic effects of Chinese herb toothpaste on fire-heat syndrome in oral cavity.</p>

Xiaoyaosan decoction regulates changes in neuropeptide y and leptin receptor in the rat arcuate nucleus after chronic immobilization stress.

The arcuate nucleus (ARC) in the basal of hypothalamus plays an important role in appetite regulation and energy balance. We sought to investigate the central neuroendocrine mechanism of appetite decrease and weight loss under chronic stress by observing the regulatory effects of Xiaoyaosan decoction in the expression of leptin receptor (ob-R) and neuropeptide Y (NPY) in the ARC. Our results showed that bodyweight and food intake of rats in the 21-day stress group increased slower than those of the normal group. Higher contents of Leptin and ob-R were noted in the 21-day stress group compared with control rats, while NPY expression was not statistically different. Xiaoyaosan powder can significantly downregulate the contents of leptin and ob-R in the hypothalamus of stressed rats. These findings suggest that increase of ob-R expression in the ARC is possibly one key central neuroendocrine change for the somatic discomfort. Weight loss and decreased food intake in rats caused by the binding of leptin to ob-R in hypothalamus do not appear to utilize the NPY pathway. This study also suggests that ob-R in the ARC may act as the target of Xiaoyaosan in regulating the symptoms such as appetite decrease and bodyweight loss under chronic stress.

[Determination of arsenic, mercury and selenium in Gynostemma pentaphyllum and rhizospheric soil samples collected from different regions by hydride generation atomic fluorescence spectrometry].

The contents of arsenic (As), mercury (Hg) and selenium (Se) in Gynostemma pentaphyllum and rhizospheric soil samples collected from seven provinces were determined, through the optimization of the hydride generation atomic fluorescence spectrometry working conditions. The results show that: the contents of As, Hg and Se in Gynostemma pentaphyllum from seven provinces revealed large differences, but compared with the limits of the two kinds of heavy metal element: As and Hg set by the Green Trade Standards of Importing & Exporting Medicinal Plants & Preparations (As < or = 2.0 mg x kg(-1), Hg < or = 0.2 mg kg(-1)), the As and Hg contents in Gynostemma pentaphyllum samples are both lower than them. The Se content in Gynostemma pentaphyllum samples and in rhizospheric soil samples revealed significant correlation, and as a result, the Gynostemma pentaphyllum from the Fu Xi area Enshi in Hu Bei province had obviously higher Se content than others in the 6 provinces. From this study, a preliminary conclusion can be drawn that Se in Gynostemma pentaphyllum is mainly from the soil, moreover, the As and Hg show the difference from Se, possibly they are still affected by the dry and wet deposition of atmospheric aerosols.

[Relation between dysbacteriosis of Helicobacter pylori and Lactobacillus acidophilus and the pathogenesis of chronic gastropathy patients of Pi-Wei damp-heat syndrome].

Pi-Wei theory is an important component of the basic theory of Chinese medicine. The pathogenesis of Pi-Wei damp-heat syndrome is an important content of Pi-Wei theory. The tongue coating is one of the most important signs reflecting Pi-Wei damp-heat syndrome. From the perspective of microecology and pathogenesis, the microbial disequilibrium caused by quantity changes of Helicobacter pylori (Hp) and Lactobacillus acidophilus (LA) and their interaction in the gastric mucosa and the tongue coating might have certain correlation with "mutual struggle between the evil and the vital qi, the disequilibrium between yin and yang". The pathogenesis features of chronic gastritis patients of Pi-Wei damp-heat syndrome was initially proposed in this article.

Relation between dysbacteriosis of Helicobacter pylori and Lactobacillus acidophilus and the pathogenesis of chronic gastropathy patients of Pi-Wei damp-heat syndrome / 中国中西医结合杂志

Pi-Wei theory is an important component of the basic theory of Chinese medicine. The pathogenesis of Pi-Wei damp-heat syndrome is an important content of Pi-Wei theory. The tongue coating is one of the most important signs reflecting Pi-Wei damp-heat syndrome. From the perspective of microecology and pathogenesis, the microbial disequilibrium caused by quantity changes of Helicobacter pylori (Hp) and Lactobacillus acidophilus (LA) and their interaction in the gastric mucosa and the tongue coating might have certain correlation with "mutual struggle between the evil and the vital qi, the disequilibrium between yin and yang". The pathogenesis features of chronic gastritis patients of Pi-Wei damp-heat syndrome was initially proposed in this article.

[Relationship between Pi-Wei damp-heat syndrome with expressions of nuclear factor-kappaB mRNA and heat shock protein 70 mRNA in patients with chronic gastritis].

OBJECTIVE: To explore the relationship between the genesis of Pi-Wei damp-heat syndrome (PDS) and the Helicobacter pylori (HP) infection in patients with chronic gastritis (CG) by observing the levels of nuclear factor kappaB (NF-kappaB) mRNA and heat shock protein 70 (HSP70) expressions. METHODS: Gastric mucous membrane tissues collected through gastroscopy from gastric antrum of 51 CG patients, 36 of PDS type (CG-PDS) and 15 of Pi-qi deficiency syndrome (CG-non-PDS) type, and 8 healthy persons (as control) were examined to diagnose the inflammation level with HE stain and to detect the existence of HP infection by rapid urease test and methylthioninium chloride stain. The mRNA expressions of NF-kappaB and HSP70 in the tissues were determined quantitatively with FQ-PCR as well. RESULTS: Patients of CG-PDS showed higher level of HSP70 mRNA expression than in the control; and level of NF-kappaB mRNA expression was higher than that in the control and CG-non-PDS (all P<0.05); but both expressions were insignificantly different in CG-PDS patients with positive or negative HP infection (P>0.05). Positive correlation of the two expressions was shown in CG-PDS with negative HP infection. CONCLUSION: NF-kappaB and HSP70 may partially embody, in some extent, the power of vital qi and evil qi in the organism; the over-expressed NF-kappaB and HSP70 may indicate the severe fighting between evil qi and vital qi, and both would be influenced to a certain degree in the fighting process.

Relationship between gene expressions of aquaporin 3 and 4 and various degrees of spleen-stomach dampness-heat syndrome in chronic superficial gastritis.

OBJECTIVE: To explore the relationship between gene expressions of aquaporin (AQP) 3 and 4 and various degrees of spleen-stomach dampness-heat syndrome in chronic superficial gastritis (CSG). METHODS: Twenty-four CSG patients were divided into three groups according to the degrees of spleen-stomach dampness-heat syndrome: mild, moderate and severe groups. There were 8 patients in each group, and another 8 healthy persons were selected as normal control. Samples of mucosa of upper stomach in the included patients and normal persons were collected by gastroscopy, and then were stored in liquid nitrogen. The gene expressions of AQP3 and AQP4 in gastric tissue were examined by fluorescence quantitative polymerase chain reaction. RESULTS: The gene expressions of AQP3 and AQP4 in the moderate and severe groups were higher than that in the mild group and normal group (P<0.05, P<0.01). The gene expression of AQP3 in the severe group was higher than that in the moderate group (P<0.05). CONCLUSION: The gene expressions of AQP3 and AQP4 in gastric mucosa are different in patients with various degrees of spleen-stomach dampness-heat syndrome of CSG. There is a relationship between expressions of AQP3 and AQP4 genes and the degrees of spleen-stomach dampness-heat syndrome, and the gene expressions will increase with the aggravation of the dampness-heat syndrome.

Relationship between Pi-Wei damp-heat syndrome with expressions of nuclear factor-kappaB mRNA and heat shock protein 70 mRNA in patients with chronic gastritis / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To explore the relationship between the genesis of Pi-Wei damp-heat syndrome (PDS) and the Helicobacter pylori (HP) infection in patients with chronic gastritis (CG) by observing the levels of nuclear factor kappaB (NF-kappaB) mRNA and heat shock protein 70 (HSP70) expressions.</p><p><b>METHODS</b>Gastric mucous membrane tissues collected through gastroscopy from gastric antrum of 51 CG patients, 36 of PDS type (CG-PDS) and 15 of Pi-qi deficiency syndrome (CG-non-PDS) type, and 8 healthy persons (as control) were examined to diagnose the inflammation level with HE stain and to detect the existence of HP infection by rapid urease test and methylthioninium chloride stain. The mRNA expressions of NF-kappaB and HSP70 in the tissues were determined quantitatively with FQ-PCR as well.</p><p><b>RESULTS</b>Patients of CG-PDS showed higher level of HSP70 mRNA expression than in the control; and level of NF-kappaB mRNA expression was higher than that in the control and CG-non-PDS (all P<0.05); but both expressions were insignificantly different in CG-PDS patients with positive or negative HP infection (P>0.05). Positive correlation of the two expressions was shown in CG-PDS with negative HP infection.</p><p><b>CONCLUSION</b>NF-kappaB and HSP70 may partially embody, in some extent, the power of vital qi and evil qi in the organism; the over-expressed NF-kappaB and HSP70 may indicate the severe fighting between evil qi and vital qi, and both would be influenced to a certain degree in the fighting process.</p>

Bioinformatics research on chronic superficial gastritis of Pi-deficiency syndrome by gene arrays.

OBJECTIVE: To determine the bioinformatical characteristics of differential gene expression in patients with chronic superficial gastritis (CSG) with the Pi-deficiency syndrome (PDS) and those of the non-Pi-deficiency syndrome (non-PDS), i.e. patients of CSG with Pi-Wei dampnese-heat syndrome and healthy persons. METHODS: With the BRB-Array Tools software package, original data collection and bioinformatic: analysis of gene arrays were conducted in 6 CSG patients of PDS (CSG-PDS), 6 CSG patients of non-PDS (CSG-nPDS), and 6 healthy volunteers (Normal). RESULTS: Compared with non-PDS, the gene expressions: in PDS with regards to protein synthesis, energy metabolism, immune reaction and ionic transport tended to be down-regulated, while those concerning secretion, cytoskeleton and ubiquitinization were up-regulated dominantly. CONCLUSIONS: The two kinds of samples, CSG-PDS/Normal and CSG-PDS/CSG-nPDS, have their respective gene expression profiles with different characteristics. Gene expression profile has certain referential significance in syndrome classification.

[The effects of safflower injection on lipid peroxidation level and expression of heme oxygenase-1 of the rat liver with hypoxia and hypercapnia].

AIM: To investigate the changes of lipid peroxidation level and expression of heme oxygenase-1 of the rat liver with chronic hypoxia and hypercapnia, and the effects of Safflower injection (a compond of Chinese Traditional medicine). METHODS: Thirty male SD rats weighing 180 approximately 220 g were divided into three groups (n=10): control group (N group), chronic hypoxia and hypercapnia for four weeks group(F group), and Safflower injection group (H group). SOD and MDA in liver tissue were measured by spectrophotometric method. And methods Immunohistochemical assay was used to detect the distribution of HO-1 protein. Pathological changes in liver tissues were observed in HE staining section. The mRNA expressions of HO-1 in liver were detected by semi-quantitative RT-PCR. RESULTS: The activity of SOD of the liver in F group were significantly lower than those in N group, and the content of MDA were significantly higher. The activity of SOD of the liver in H group were significantly higher than those in F group, and the content of MDA were significantly lower. In F group there were multiple dispersed immunoreactivity cells in liver. And compared to those in F group, the immunoreactivity cells were significantly decreased in H group. HE staining revealed that there were many hepatocytes with obvious adipose degeneration. Hepatic pathological damage in H group was slighter than that in F group. The expression of HO-1 mRNA of the liver in F group were significantly higher than those in N group (P < 0.01), and those in H group were significantly lower than those in F group (P < 0.01) . CONCLUSION: Chronic hypoxia and hypercapnia increases the level of oxidative stress. Safflower injection have a protective effect, maybe because of the accommodation of the expression of HO-1 of the liver and the elimination of free radicals.